Mycobacterium avium is a major opportunistic pathogen in AIDS patients as well as a major cause of pulmonary infection in individuals with underlying lung disease. However, our understanding of M. avium pathogenesis is lacking due in part to limited information on M. avium components involved in modulating the host response. Nevertheless, previous studies suggest that glycopeptidolipids (GPLs), which are major surface components expressed on many non-tuberculosis mycobacteria, may function in directing the host response to an M. avium infection. We have found that GPLs can engage the pattern recognition receptor Toll-like receptor 2 (TLR2) leading to macrophage activation. Interestingly, we found that GPLs varied in their ability to interact with TLR2 and that slight changes in the GPL acetylation and methylation patterns determine whether this glycolipid can signal through TLR2. GPLs can also engage the mannose receptor and that this interaction is required for the delayed accusation of lysosomal markers observed for phagosomes containing GPL-coated beads. Based on these observations we hypothesize that M. avium can modify its GPL structure during the course of an infection and that this is an important virulence mechanism. Further, we predict that increased expression of certain GPL variants by M. avium will correlate with a strain's pathogenicity. To test these predictions we will: 1) Define the GPL structural components necessary for its binding to the MR and for its ability to delay phagosome maturation and the mechanism by which GPLs mediate this delay. 2) Characterize the GPL composition of M. avium strains following macrophage and mouse infections and define how the GPL composition correlates with strain virulence. 3) Generate M. avium knockout mutants for genes involved in GPL biosynthesis and evaluate the mutants for GPL-mediated activities including signaling through TLR2 and MR, biofilm formation and sliding motility and for virulence in a mouse infection model. Upon completion of these studies we will have a better understanding of M. avium pathogenesis and the role that GPLs play in this process and the information garnered may lead to the development of new diagnostic tools to evaluate M. avium virulence. PUBLIC HEALTH RELEVANCE: Mycobacterium avium is a major opportunistic pathogen in AIDS patients and a significant cause of morbidity and mortality in HIV infected individuals. It is also a common cause of pulmonary infections in individuals with underlying lung disease. The goal of this proposal is to better understand M. avium pathogenesis so as to develop new methods to limit M. avium infections and to treat those infected.